Hlola ukusebenzisana kwamabhayisikili ashisayo ocwaningweni

Amabhayisikili ashisayo, awaziwa nangokuthi imishini ye-PCR, angamathuluzi abalulekile kubhayoloji yamangqamuzana nocwaningo lwezofuzo. Lezi zinsimbi zisetshenziselwa ukukhulisa i-DNA ne-RNA ngobuchwepheshe be-polymerase chain reaction (PCR). Kodwa-ke, ukuguquguquka kwamabhayisikili ashisayo akukhawulelwe ezinhlelweni ze-PCR. Kulesi sihloko, sizohlola izindlela ezehlukene zamabhayisikili ashisayo asetshenziswa ocwaningweni kanye nokubaluleka kwawo ekuthuthukiseni ulwazi lwesayensi.

1. Ukukhulisa i-PCR

Umsebenzi oyinhloko we-aumjikelezo oshisayoukwenza i-PCR amplification, okubalulekile ezinhlobonhlobo zezinhlelo zokusebenza zebhayoloji yamangqamuzana. Ngokubeka ngaphansi kwesampula ye-DNA noma ye-RNA ochungechungeni lwezinguquko zezinga lokushisa, amabhayisikili ashisayo akhuthaza ukukhishwa kwe-denaturation, annealing, kanye nokwelulwa kwemicu ye-nucleic acid, okuholela ekukhuliseni okucacile kokulandelana okuqondiwe okuqondiwe. Le nqubo ibalulekile ekuhlaziyweni kofuzo, izifundo zesisho sezakhi zofuzo, kanye nokutholwa kwezinto ezithathelwanayo.

2. I-Quantitative PCR (qPCR)

Ngokungeziwe ku-PCR ejwayelekile, amabhayisikili ashisayo asetshenziselwa ubuningi be-PCR noma i-qPCR, okuvumela ukulinganisa okuhlosiwe kwe-nucleic acid kusampula. Ngokuhlanganisa odayi be-fluorescent noma ama-probe, amabhayisikili ashisayo angakwazi ukukala ukuqoqwa kwemikhiqizo ye-PCR ngesikhathi sangempela, anikeze imininingwane ebalulekile emazingeni okubonisa izakhi zofuzo, umthamo wegciwane egazini, nokuhlukahluka kofuzo.

3. Reverse transcription PCR (RT-PCR)

Amabhayisikili ashisayo adlala indima ebalulekile ekubhalweni okuhlanekezelwe kwe-PCR, indlela eguqula i-RNA ibe i-DNA ehambisanayo (cDNA) ukuze kukhuliswe okulandelayo. Le ndlela ibalulekile ekutadisheni ukubonakaliswa kofuzo, amagciwane e-RNA, namaphethini okuhlangana kwe-mRNA. Ibhayisikili elishisayo elinokulawula okunembayo kwezinga lokushisa libalulekile empumelelweni yokuhlolwa kwe-RT-PCR.

4. I-PCR yedijithali

Intuthuko kubuchwepheshe be-thermal cycler iholele ekuthuthukisweni kwe-PCR yedijithali, indlela ebucayi kakhulu yokulinganisa okuphelele kwama-nucleic acid. Ngokuhlukanisa ukusabela kwe-PCR kube izinkulungwane zama-microreactions angawodwana, amabhayisikili ashisayo angakwazi ukunquma ngokunembile ukuhlangana kokuqala kwe-molecule eqondiwe, okwenza i-PCR yedijithali ibe ithuluzi elibalulekile lokutholwa kokuguqulwa kwezakhi ezingavamile kanye nokuhlaziya ukuhlukahluka kwenombolo.

5. Ukulungiswa kwemitapo yolwazi elandelanayo yesizukulwane esilandelayo

Amabhayisikili ashisayo ayingxenye ebalulekile yenqubo yokulungiselela umtapo wezinhlelo zokusebenza zokulandelana kwesizukulwane esilandelayo (NGS). Ngokwenza ukukhulisa okusekelwe ku-PCR kwezingcezu ze-DNA, amabhayisikili ashisayo anika amandla ukwakhiwa kwemitapo yolwazi elandelanayo kusukela ezintweni zokuqala ezilinganiselwe, okuvumela abacwaningi ukuthi bahlaziye lonke i-genome, i-transcriptome, noma i-epigenome ephilayo.

6. Ubunjiniyela bamaprotheni kanye ne-Mutagenesis

Ngaphezu kwe-nucleic acid amplification, amabhayisikili ashisayo asetshenziselwa ubunjiniyela bamaprotheni kanye nezifundo ze-mutagenesis. I-mutagenesis eqondiswe kusayithi, ukwenziwa ngcono kwe-protein expression, kanye nokuhlolwa kokuziphendukela kwemvelo okuqondisiwe kuvame ukuncika kumasu asekelwe ku-PCR, futhi amabhayisikili ashisayo anokulawula izinga lokushisa okunembile kanye namazinga okufudumeza afanayo kanye nokupholisa kubalulekile ukuze kutholwe imiphumela enembile nekwazi ukukhiqizwa kabusha.

7. Ukuhlola ukuphepha kwemvelo nokudla

Amabhayisikili ashisayo nawo asetshenziswa ekuhloleni ukuphepha kwemvelo nokudla, ikakhulukazi ukutholwa kwamagciwane amancane, i-genetically modified organisms (GMOs) kanye nama-pathogens atholakala ekudleni. Ukuhlolwa okususelwe ku-PCR kwenziwa kumabhayisikili ashisayo kunika amandla ukuhlonza ukungcola okusheshayo nokuqondile, okuqinisekisa ukuphepha kanye nekhwalithi yamasampuli okudla nawemvelo.

Ngokufigqiwe,amabhayisikili ashisayozingamathuluzi abalulekile kubhayoloji yamangqamuzana nocwaningo lwezakhi zofuzo, ahlinzeka ngezinhlelo eziningi zokusebenza ngale kokukhulisa i-PCR yendabuko. Ukusebenziseka kwazo ngezindlela ezihlukahlukene nokunemba kwazo kuzenza zibaluleke kakhulu ekuhloleni kusukela ekuhlaziyweni kwenkulumo yofuzo kuye ekuqaphelweni kwemvelo. Njengoba ubuchwepheshe buqhubeka nokuthuthuka, amabhayisikili ashisayo kungenzeka adlale indima ebaluleke kakhulu ekuqhubekiseni ukutholwa kwesayensi nokusungula izinto ezintsha.